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[共表达乙肝表面抗原和粒细胞巨噬细胞集落刺激因子的质粒的抗病毒作用]

[Antivirus effect of plasmid co expressing hepatitis B surface antigen and granulocyte macrophage-colony stimulating factor].

作者信息

Qing Yu-ling, Zhao Jia-jiang, Ren Hong

机构信息

Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2004 Apr;12(4):201-4.

PMID:15099465
Abstract

OBJECTIVE

To investigate whether GM-CSF can enhance the antiviral effect of HBsAg DNA vaccine.

METHODS

Divided animals into 8 groups. Group A: pcDNA3.1-S 100microg; Group B: pcDNA3.1-GM-CSF-S 100microg; Group C: pcDNA3.1-S-GM-CSF 100microg; Group D: pcDNA3.1-S 50microg + pcDNA3.1-GM-CSF 50microg; Group E: pcDNA3.1-GM-CSF 100microg; Group F: recombinant HBsAg vaccine 1microg; Group G: pcDNA3.1,100microg; Group H: PBS 100microl. Serum HBsAg level and concentration of IL-2, IL-4 and IFN-gamma were examined using commercial ELISA kit. The [3H] thymidine incorporation into DNA of Spleen cells was measured; HBsAg expression of hepatocytes from HBV-transgenic mice was assessed using immunohistochemical analysis.

RESULTS

Serum HBsAg level was lower and concentration of IL-2, IFN-gamma and SI was higher in mice immunized with pcDNA3.1-GM-CSF-S than those from mice immunized with pcDNA3.1-S and other groups (F=11.262, P<0.01, F=8.147, P<0.01, F=62.275, P<0.01, F=116.28, P<0.01. Less Hepatic HBsAg expression and decline of pcDNA3.1-GM-CSF-S of mice immunized with pcDNA3 were observed in comparison with control groups (F=41.439, P<0.01). Liver histological analysis showed no evidence of necrosis or inflammation in various groups.

CONCLUSION

The plasmid co expressing GM-CSF and HBsAg could improve HBsAg-specific humoral and cellular immune responses induced by plasmid encoding HBsAg alone and enhance HBsAg DNA vaccine antivirus effect.

摘要

目的

研究粒细胞-巨噬细胞集落刺激因子(GM-CSF)是否能增强乙肝表面抗原(HBsAg)DNA疫苗的抗病毒效果。

方法

将动物分为8组。A组:pcDNA3.1-S 100微克;B组:pcDNA3.1-GM-CSF-S 100微克;C组:pcDNA3.1-S-GM-CSF 100微克;D组:pcDNA3.1-S 50微克 + pcDNA3.1-GM-CSF 50微克;E组:pcDNA3.1-GM-CSF 100微克;F组:重组HBsAg疫苗1微克;G组:pcDNA3.1,100微克;H组:PBS 100微升。使用商用酶联免疫吸附测定(ELISA)试剂盒检测血清HBsAg水平以及白细胞介素-2(IL-2)、白细胞介素-4(IL-4)和γ-干扰素(IFN-γ)的浓度。测定[3H]胸腺嘧啶核苷掺入脾细胞DNA的情况;采用免疫组织化学分析评估乙肝病毒(HBV)转基因小鼠肝细胞的HBsAg表达。

结果

用pcDNA3.1-GM-CSF-S免疫的小鼠血清HBsAg水平低于用pcDNA3.1-S免疫的小鼠及其他组,IL-2、IFN-γ和SI的浓度高于用pcDNA3.1-S免疫的小鼠及其他组(F = 11.262,P < 0.01;F = 8.147,P < 0.01;F = 62.275,P < 0.01;F = 116.28,P < 0.01)。与对照组相比,用pcDNA3.1-GM-CSF-S免疫的小鼠肝组织中HBsAg表达减少且pcDNA3.1-GM-CSF-S下降(F = 41.439,P < 0.01)。肝脏组织学分析显示各实验组均无坏死或炎症迹象。

结论

共表达GM-CSF和HBsAg的质粒能增强单独编码HBsAg的质粒诱导的HBsAg特异性体液免疫和细胞免疫反应,并增强HBsAg DNA疫苗的抗病毒效果。

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